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Chemiluminescent enzyme-linked immunoassay detection kit for staphylococcal enterotoxin C

A staphylococcal intestinal and chemiluminescent enzyme technology, applied in the field of immunoassay, can solve the problems of poor detection sensitivity, many interference factors, low sensitivity, etc., and achieve the effects of strong specificity, good repeatability and high sensitivity

Active Publication Date: 2013-02-27
FOURTH MILITARY MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The disadvantages of immunoagar diffusion method and reverse indirect hemagglutination method are time-consuming, many interference factors affecting the analysis, low sensitivity and poor specificity
The immunochip method is expensive, operators need to undergo professional training, there are many interference factors affecting the analysis, and the sensitivity is low.
Although enzyme-linked immunosorbent assay is easy to operate and suitable for large-throughput detection, its detection sensitivity is poor

Method used

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  • Chemiluminescent enzyme-linked immunoassay detection kit for staphylococcal enterotoxin C
  • Chemiluminescent enzyme-linked immunoassay detection kit for staphylococcal enterotoxin C
  • Chemiluminescent enzyme-linked immunoassay detection kit for staphylococcal enterotoxin C

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Example 1: Preparation of Staphylococcus aureus Enterotoxin C Monoclonal Antibody and Screening of Coating Antibody and Detection Antibody Pairing

[0046] (1) Preparation of anti-Staphylococcus aureus enterotoxin C monoclonal antibody: The purified natural Staphylococcus aureus enterotoxin C antigen standard was provided by the Institute of Microbiology and Epidemiology, Academy of Military Medical Sciences. Eight-week-old BALB / c mice were used as immunized animals, with Staphylococcus aureus enterotoxin C as the immunogen, the first immunization dose was 50 μg / mL, and the immunogen was dissolved in normal saline and an equal volume of Freund’s complete adjuvant Emulsifier, multi-point subcutaneous injection. The second immunization was carried out after an interval of 3 weeks. The dose and route were the same as the first immunization, and the adjuvant was Freund's incomplete adjuvant. After 2-3 weeks, the third immunization was carried out, the dose was the same as ...

Embodiment 2

[0053] Example 2: Establishment of Chemiluminescent ELISA Method

[0054] (1) Selection of optimal coating antibody concentration:

[0055] The coating antibody was coated on the chemiluminescent enzyme-linked plate at serial dilutions of 1.25 μg / mL, 2.5 μg / mL, 5 μg / mL, 10 μg / mL, 20 μg / mL and 40 μg / mL, 100 μL / well, incubated overnight at 4 °C, Wash the plate 3 times with washing solution; block with 250 μL / well blocking solution, place at room temperature for 1 hour, and wash the plate 3 times; add 0.01 ng / mL Staphylococcus aureus enterotoxin C standard antigen, 100 μL / well, and each coating The antibody concentration was used as blank control wells, incubated at 37°C for 1 hour, and washed the plate 3 times; respectively added 1:16000 diluted anti-Staphylococcus aureus enterotoxin C detection antibody, incubated at 37°C for 1 hour, washed the plate 5 times; added 100 μL / well of chemiluminescence solution, and measure the luminescence value. According to the ratio of the me...

Embodiment 3

[0065] Example 3: Application of chemiluminescent ELISA detection kit for detection of staphylococcal enterotoxin C in different matrices

[0066] This example is used to evaluate the accuracy and practicability of the staphylococcal enterotoxin C chemiluminescence ELISA detection kit of the present invention in detecting staphylococcus aureus enterotoxin C in different matrices.

[0067]The standard substance of Staphylococcus aureus enterotoxin C was diluted to different concentrations with environmental matrix river water, food matrix milk, body fluid matrix human serum or human urine, and the content of Staphylococcus aureus enterotoxin C was detected by chemiluminescence enzyme immunoassay, and calculated The ratio of the measured concentration to the actual concentration added is the recovery rate.

[0068] (1) Pre-coat the chemiluminescence enzyme-linked plate with the coating antibody: dilute the coating antibody to 2.5 μg / mL with the coating solution and add it to the...

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Abstract

The invention discloses a chemiluminescent enzyme-linked immunoassay detection kit for staphylococcal enterotoxin C. The contents of the kit comprise a staphylococcal enterotoxin C-resistant monoclonal antibody, a horseradish peroxidase-marked staphylococcal enterotoxin C-resistant detection antibody solution, a staphylococcal enterotoxin C series standard solution, a coating solution, a sealing solution, a cleaning solution, a chemiluminescent solution and a chemiluminescent enzyme-linked plate. The kit has the characteristics of high specificity, sensitivity and repeatability, and is suitable for monitoring diseases and quickly and quantitatively detecting staphylococcal enterotoxin C in foods and blood.

Description

technical field [0001] The invention belongs to the technical field of immune analysis, and relates to a kit for detecting superantigens, in particular to a staphylococcal enterotoxin C chemiluminescent ELISA detection kit, which uses a double-antibody sandwich chemiluminescent ELISA This method is used to quantitatively detect the content of Staphylococcus aureus enterotoxin C in food, especially in milk. Background technique [0002] Food safety is closely related to human health, and food safety has become a hot issue of social concern. With the help of the development of immunology technology, building a complete foodborne disease monitoring system and implementing all-round food safety monitoring will help to improve the product quality of the food industry and thus protect the health of the public. [0003] Staphylococcus aureus is an important pathogenic bacterium of zoonosis, which widely exists in air, water, dust, skin and nasal cavity of warm-blooded animals such...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N21/76
Inventor 李琦刘志佳黄业青杨琨张春梅宋朝君李永明徐竹蔚刘飞陈丽华张赟方亮孙元杰易静周幸春马樱刘蓓张宇丝刘蓉蓉金伯泉
Owner FOURTH MILITARY MEDICAL UNIVERSITY
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